Anti-aging composition comprising a plant extract

ABSTRACT

The present description relates to an anti-aging composition comprising at least one plant extract. The composition can comprise at least two different anti-aging agents, wherein one the anti-aging agent is a plant extract. The combination of two anti-aging agents has a superior effect on regulating longevity compared to the use of one anti-aging agent. The plant extract is preferably selected from the group consisting of a Black cohosh extract, a Valerian extract, a  Ginkgo biloba  extract, a Celery seed extract, a White willow extract and a Passion flower extract.

TECHNICAL FIELD

The present description relates to an anti-aging composition comprisingat least one plant extract.

BACKGROUND ART

It is known that aging drives disease. Nearly every major killer diseasein developed countries shares a common feature: the risk of getting thedisease increases dramatically as you get older. For example, thelikelihood of being diagnosed with Alzheimer's disease doubles everyfive years after the age of 65. A similar kind of relationship can beseen for most types of cancer, heart disease, diabetes, kidney disease,and many others.

The rate of aging may also be measured, and an accelerated rate of agingmay be considered ‘premature aging’, while a slower rate of the agingprocess may extend health span. It is desirable to maximize the healthylifespan of cells and organisms and it is also desirable to extend thehealthy lifespan by decreasing the rate of aging process and the onsetof dysfunctional or disease states. Shortening the lifespan and/oraccelerating apoptosis of unhealthy, diseased, damaged, or cancerouscells may also be desirable.

Rather than focussing on curing the individual disease, interventionsthat target the molecular processes causing aging can simultaneouslydelay the onset and progression of most age-related disorders (Longo etal., 2015, Aging Cell, 1-14). Such interventions are predicted to have agreater beneficial effect on healthy lifespan than the one that can beattained by treating individual diseases.

Recent discoveries suggest that aging is neither driven by accumulationof molecular damage of any cause, nor by random damage of any kind.Studies in humans and model organisms aimed at elucidating the molecularmechanisms of aging have demonstrated the existence of broadly conservedlongevity pathways, and, for the first time, offer real hope ofintervening to enhance healthy aging. The best-characterizedintervention for delaying aging is dietary restriction (also referred toas caloric restriction). Many studies have shown that a reduced calorieregimen can increase lifespan and delay the onset of multipleage-related phenotypes in a diverse range of organisms, including theentire major model systems used in biomedical research.

Not surprisingly, dietary restriction modulates the activity of multiplecellular factors, several of which have been implicated in longevity andhealth span. These factors include sirtuins, key metabolic regulatorssuch as AMP kinase, antioxidant enzymes, DNA damage response enzymes,and others. Among these, however, the mTOR signaling pathway, inparticular, has emerged as a central pro-aging pathway that is inhibiteddue to pro-longevity effects of dietary restriction in yeast, nematodesand fruit flies. In response to nutrient depletion, mTOR activity isreduced and this results in a cascade of downstream events that havebeen shown to promote longevity and enhance resistance to stress. Inparticular, reduced synthesis of new proteins via inhibition of mRNAtranslation, enhanced degradation of damaged proteins and othermacromolecules via autophagy, and altered carbon metabolism andmitochondrial function all contribute to lifespan extension by dietaryrestriction.

Although caloric restriction can provide significant benefits, itsimplementation in humans is unlikely to be achieved. Indeed, toimplement such dietary regimen for an adult individual, such individualmust limit food consumption to the equivalent of a 5 year old child whois not too active.

Other than dietary restriction, the only non-genetic intervention knownto exhibit a significant lifespan-extending effect in yeast, nematodes,fruit flies, and mice is the mTOR-inhibiting drug called rapamycin.However, rapamycin have many negative side effects on an organism, andnatural products that inhibit mTOR but lack such negative side effectsremain to be identified. Further, dietary restriction is known to haveeffect not only on mTOR, but also on the AMPK, sirtuins and insulinsignaling pathways.

Therefore, the implementation of complex mixtures of several (or many)natural products modulating different signaling pathways is desirable toachieve a more important anti-aging effect.

Classic broad symptoms of aging in mammalian species include increasedcurvature of the spine (kyphosis), reduced fertility, loss of hearingand eyesight, graying and loss of hair, anemia and immune failure,weight loss, frailty, and loss of cognition. These systemic changes aredriven by a variety of molecular, biochemical, and metabolic alterationsthat occur at the cellular level. A particularly important outcome forcellular aging studies was the use of yeast to discover the conservedgenetic pathways that modulate longevity across broad evolutionarydistance. Chronological lifespan of yeast cells in stationary culture isthe most fruitful model in aging research and numerous papers coveringthis topic have been published (see for example Kaeberlein, 2010,Nature. 2010 Mar. 25; 464(7288): 513-519).

Humans have evolved to have significantly extended (as compare to mostof mammalian species) longevity. Thus, any additional gains in maximumlifespan are likely to be minimal; however, interventions thatsignificantly extend lifespan in model organisms have the potential toextend health span in humans and, therefore, to cause a substantialreduction in morbidity.

The molecular, cellular, organismal and genetic mechanisms that controlaging and lifespan have been shown to be highly conserved acrossmillions of years of evolution. Therefore, responses of lower eukaryoticorganisms (e.g., C. elegans, D. melanogaster, S. cerevisiae) to geneticand pharmacological interventions extending longevity are expected to besimilar in mammals including humans. Thus, there is an urgent need inidentifying pharmaceutical compositions that mimic aging-delayingeffects of dietary/caloric restriction or lifespan-extending geneticmutations.

The budding yeast, S. cerevisiae, has been used extensively as a modelfor cellular aging (Kaeberlein, 2010, Nature, 25: 513-519).Chronological lifespan in yeast is similar to aging of post-mitoticcells, such as mature neurons, adipocytes and mature muscle cells. Yeastchronological aging can be therefore compared to and is predictive ofaging of cells and tissues in a human organism. Chronological aging inyeast is assessed by growing a culture of cells to maximal density, atwhich point nutrients become limiting and cell division arrests. Afraction of cells that can re-enter the cell cycle when exposed tonutrient-rich media is considered to be a fraction of viable cells, andindividual cultures are followed until cell viability is close to zero.

While treatments exist for some symptoms of aging-associated disorders,no treatments are currently known that delay aging of the entireorganism by targeting multiple cellular and organismal processes withthe help of natural extract(s). In addition, by slowing the rate ofaging, it may be possible to delay the onset of variousdiseases/conditions associated with aging.

There is thus still a need to be provided with an anti-aging compositionfor delaying aging.

SUMMARY

In accordance with the present description there is now provided ananti-aging composition comprising at least two different anti-agingagents, wherein said at least two anti-aging agents are two differentplant extracts or one plant extract and a second anti-aging agent, andwherein the at least two anti-aging agents have an superior effect onregulating longevity compared to one anti-aging agent.

It is thus provided an anti-aging composition comprising at least twodifferent anti-aging agents, wherein the at least two anti-aging agentsare two different plant extracts or one plant extract and a secondanti-aging agent, wherein the plant extract is at least one of a Blackcohosh extract, a Valerian extract, a Ginkgo biloba extract, a Celeryseed extract, a White willow extract and a Passion flower extract, andwherein the at least two anti-aging agents have an superior effect onregulating longevity compared to one anti-aging agent.

It is also provided an anti-aging composition comprising at least oneplant extract selected from the group consisting of a Black cohoshextract, a Valerian extract, a Ginkgo biloba extract, a Celery seedextract, a White willow extract and a Passion flower extract, and acarrier.

In an embodiment, the second anti-aging agent is resveratrol, metformin,myriocin, or spermidine.

In another embodiment, the plant extract is selected from the groupconsisting of a Black cohosh extract, a Valerian extract, a Ginkgobiloba extract, a Celery seed extract, a White willow extract and aPassion flower extract.

In a further embodiment, the at least two anti-aging agents modulate atleast two pathways that regulate longevity.

In a supplemental embodiment, the pathways are TORC1, cAMP/PKA, PKH1/2,SNF1/AMPK or ATG pathways.

In an embodiment, the composition comprises a combination of a Blackcohosh extract, and/or a Valerian extract, and/or a Ginkgo bilobaextract, and/or a Celery seed extract, and/or a White willow extract,and/or Passion flower extract.

In an embodiment, the composition comprises a combination of a Blackcohosh extract with a Valerian extract, a Ginkgo biloba extract, aCelery seed extract, a White willow extract or Passion flower extract.

In another embodiment, the composition comprises a combination of aValerian extract with a Ginkgo biloba extract, a Celery seed extract orPassion flower extract.

In another embodiment, the composition comprises a combination of aPassion flower extract with a Ginkgo biloba extract or a Celery seedextract.

In another embodiment, the composition comprises a combination of aGinkgo biloba extract and a Celery seed extract.

In another embodiment, the composition described herein furthercomprises resveratrol, myriocin, metformin, or spermidine.

In an embodiment, the composition comprises a Black Cohosh extractand/or Spermidine.

In an embodiment, the composition comprises a Valerian extract withResveratrol, metformin, myriocin and/or Spermidine.

In another embodiment, the composition comprises a Ginkgo biloba extractand myriocin and/or Spermidine.

In another embodiment, the composition comprises a passion flowerextract and resveratrol, metformin, myriocin, and/or spermidine.

In another embodiment, the composition comprises a Celery seed extractand Resveratrol, metformin, and/or myriocin.

In an embodiment, the composition comprises a White willow extract andmetformin, myriocin, and/or spermidine.

In a further embodiment, the composition described herein delay theonset and progression of age-related disorders.

In another embodiment, the age-relate disorders are cardiovasculardisorders, glycemic disorders, neurodegenerative disorders orosteoporosis disorders.

In another embodiment, the composition described herein is formulated asa cosmetic composition, a dermatological composition, a nutraceuticalcomposition or a pharmaceutical composition.

It is also provided the use of the composition described herein forprolonging longevity of a subject.

It is also provided a method of prolonging longevity of a subjectcomprising administering to the subject an effective amount of thecomposition described herein.

BRIEF DESCRIPTION OF THE DRAWINGS

Reference will now be made to the accompanying drawings.

FIG. 1 illustrates mean and maximum lifespan evaluation.

FIG. 2 illustrates a null effect of extracts 36 and 16 on the lifespanof S. cerevisiae BY4742.

FIG. 3 illustrates a schematic representation of Black cohosh extract(PE4), Valerian extract (PE5), Passion flower extract (PE6), Ginkgobiloba extract (PE8), Celery seed extract (PE12) and White willowextract (PE21) delaying yeast chronological aging and have differenteffects on several longevity defining cellular processes. Arrowspointing at boxes with the terms of longevity-defining cellularprocesses denote activation of these processes, T bars denote inhibitionof these processes, whereas lines with filled circles denote change inthe age-related chronology of intracellular ROS. The thickness of sucharrows, T bars and lines with filled circles correlates with the extentto which a PE activates, inhibits or alters the age-related chronology(respectively) of a particular longevity-defining cellular process.Arrows and T bars pointing at boxes with the term “AGING” denoteacceleration or deceleration (respectively) of yeast chronologicalaging.

FIG. 4 illustrates how the Black cohosh extract (PE4), Valerian extract(PE5), Passion flower extract (PE6), Ginkgo biloba extract (PE8), Celeryseed extract (PE12) and White willow extract (PE21) delay yeastchronological aging via the longevity-defining network of signalingpathways/protein kinases. Activation arrows and inhibition bars denotepro-aging processes or anti-aging processes.

FIG. 5 illustrates the delaying of aging process by the synergy of aPassion flower extract (NP#6) with resveratrol.

FIG. 6 illustrates the increase in cellular concentrations ofphosphatidic acid (PA), phosphatidylserine (PS),phosphatidylethanolamine (PE), phosphatidylcholine (PC) andphosphatidylinositol (PI), the decrease in cellular concentration oftriacylglycerols in chronologically aging yeast treated with the WhiteWillow extract (NHP #21).

DETAILED DESCRIPTION

It is provided an anti-aging composition comprising at least one plantextract.

It is also provided an anti-aging composition comprising at least twodifferent anti-aging agents. The anti-aging agents are at least twoplant extracts or a combination of one plant extract and a second agent,being for example resveratrol or spermidine. The composition comprisingtwo agents has a superior effect on regulating longevity compared to theuse of each individual anti-aging agent.

As described herein, the plant extract is preferably selected from thegroup consisting of a Black cohosh extract, a Valerian extract, a Ginkgobiloba extract, a Celery seed extract, a White willow extract and aPassion flower extract.

In another embodiment, the present disclosure describes the use of atleast two different anti-aging agents able to modulate at least twodifferent pathways that regulate longevity to obtain the most importantanti-aging effect, said pathways being for example TORC1, cAMP/PKA,PKH1/2, SNF1/AMPK and ATG pathways.

More particularly, it is provided an anti-aging composition comprisingat least one plant extract selected from the group consisting of a Blackcohosh extract, a Valerian extract, a Ginkgo biloba extract, a Celeryseed extract, a White willow extract and a Passion flower extract.

A method and composition for delaying the onset of aging process isprovided. Described herein are methods and compositions for alteringmitochondrial biogenesis and/or mitochondrial maintenance, respiratoryefficiency, DNA maintenance, DNA repair, gene expression, and/or genefunction, for instance in order to (in various embodiments) reduceand/or retard rate of senescence of a cell, tissue, organ, and/ororganism. In example embodiments, this involves altering the maintenanceor function of telomeres and telomere structure, the maintenance andcontrol of the cellular responses to oxidative stress and/or oxidativeDNA damage, and cellular response to environmental damage or disease orimmune response or genetic alteration of cells (see FIG. 3). Morespecifically, the present description: (i) relates generally to thefield of aging process; and (ii) describes novel compositions andmethods for using plant extracts or pure chemical compounds, alone or inmixtures and having therapeutic uses in mammals

The present description relates to the cosmetic, dermatological,nutraceutical or pharmaceutical (therapeutic) use of natural compounds,in particular as agents that enable prolongation of the lifespan of acell, i.e. compounds acting as potent anti-aging agents.

The present disclosure relates generally to compounds and methods thatenhance survivability and treat and protect cells and animals frominjury, disease, and premature death. The composition described hereinmaximizes the healthy lifespan and/or extend health span. Accordingly,it is encompassed a composition that delay the onset and progression ofmost age-related disorders, such as for example cardiovasculardisorders, glycemic disorders, neurodegenerative disorders orosteoporosis disorders.

A total of 59 plant extracts were selected as the one that maypotentially have anti-aging effects. FIG. 2 demonstrates examples of‘negative’ extracts. Among the 59 plant extracts tested, 6 extracts werefound to be positives. More than 10 000 trials were performed toidentify such positive extracts, to establish their most efficientlongevity-extending concentrations and to demonstrate the beneficialeffects in combining them. The extraction process and the commercialsource are not limiting factors for the potency of the anti-aging plantextracts described herein.

Data presented herein demonstrates that different anti-aging plantextracts delay aging by targeting different signaling pathways oflongevity regulation and/or individual protein components of suchpathways. Each of these anti-aging extracts, as well as well-knownanti-aging chemical compounds, influences the TORC1, cAMP/PKA, PKH1/2,SNF1/AMPK and ATG pathways that regulate longevity (see FIG. 4). It isthus demonstrated that:

-   -   (i) Black cohosh extract (NP #4) delays aging by attenuating the        inhibiting effect of the TORC1 signaling pathway on the        AMP-activated protein kinase SNF1/AMPK;    -   (ii) Valerian extract (NP #5) delays aging by mitigating two        arms of the pro-aging cAMP/PKA signaling pathway related to the        Gh/IGF-1 axis;    -   (iii) Ginkgo biloba extract (NP #8) delays aging by weakening        the inhibiting effect of the pro-aging cAMP/PKA signaling        pathway on the AMP-activated protein kinase SNF1/AMPK;    -   (iv) Celery seed extract (NP #12) delays aging by activating the        nutrient-sensing protein kinase Rim15, on which the pro-aging        TORC1 and cAMP/PKA signaling pathways converge;    -   (v) White willow extract (NP #21) delays aging by attenuating        SCH9, a nutrient-sensory protein kinase known to be activated by        the pro-aging TORC1 and PKH1/2 signaling pathways; and    -   (vi) Passion flower extract (NP #6) delays aging not by        targeting currently known pro- or anti-aging pathways of        longevity regulation; thus, this plant extract inhibits a        presently unknown pro-aging pathway and/or activates a presently        unknown anti-aging pathway.

The existence of such mechanism for modulating the TORC1, cAMP/PKA,PKH1/2, SNF1/AMPK and ATG pathways by different anti-aging plantextracts as well as by well-known anti-aging chemical compounds impliesthat multi-component mixtures of some previously unknown anti-agingplant extracts and certain known anti-aging compounds will concomitantlyattenuate several pro-aging signaling pathways and activate severalanti-aging signaling pathways. Thus, such mixtures should delay agingprocess to a significantly higher extent than any previously knowndietary interventions.

The pairwise mixtures of the many plant extracts described hereinextended the mean and maximum lifespans of chronologically aging yeastto significantly higher extent that each of the plant extracts alone.

Aging of multicellular and unicellular eukaryotic organisms is a complexbiological phenomenon affecting many cellular processes. These numerouscellular processes are modulated by signaling pathways that areconserved across phyla and include the insulin/insulin-like growthfactor 1 (IGF-1), AMP-activated protein kinase/target of rapamycin(AMPK/TOR) and cAMP/protein kinase A (cAMP/PKA) pathways. In yeast,worms, fruit flies and mammals these signaling pathways converge into anetwork regulating aging process. This network responds to theage-related partial mitochondrial dysfunction and is modulated bymitochondrially produced reactive oxygen species (ROS). By sensing thenutritional status of the whole organism as well as the intracellularnutrient and energy status, functional state of mitochondria, andconcentration of ROS produced in mitochondria, the aging process networkregulates lifespan and healthspan across species.

In yeast, network regulating longevity includes the following signalingpathways: (1) the pro-aging PKA (protein kinase A) pathway; (2) thepro-aging TORC1 (target of rapamycin complex 1) pathway; (3) thepro-aging PKH1/2 (Pkb-activating kinase homolog) pathway; (4) theanti-aging SNF1 (sucrose non-fermenting) pathway; (5) the anti-aging ATG(autophagy) pathway. Moreover, SCH9 is a pro-aging protein kinasestimulated by the TORC1 and PKH1/2 pathways, whereas RIM15 is ananti-aging protein kinase inhibited by the PKA and TORC1 pathways.

Each of the plant extracts exhibiting an extremely high anti-agingefficiency (as compare to the dietary restriction impact) was added tothe different mutant cultures, at a concentration that was found to beoptimal for its longevity-extending action.

These experiments revealed that certain plant extracts greatly delayaging by inhibiting only the pro-aging TOR signaling pathway, some plantextracts extend longevity by attenuating only the pro-aging cAMP/PKAsignaling pathway, certain plant extracts delay aging by mitigating boththe TOR and cAMP/PKA pathways, whereas some plant extracts extendlongevity by targeting cellular processes that are not orchestrated byany of these two pro-aging signaling pathways (FIG. 4).

Because passion flower extract (NP #6) and resveratrol target differentpro- and anti-aging signaling pathways, their mixtures were expected toexhibit synergistic anti-aging effects. Indeed, a mixture of NP #6 andresveratrol exhibits a synergistic extending effect on longevity ofyeast cells (FIG. 5). As this inhibitory effect on aging process is themost important ever seen, the analysis of cell lipids was done to have abetter appreciation of this synergy.

The results of this experiment imply that Passion flower extract (NP #6)alone and together with resveratrol:

-   -   i) Significantly increases the level of cardiolipin (this lipid        can be found only in mitochondria), phosphatidylcholine,        phosphatidylethanolamine, phosphatidylinositol;    -   ii) Significantly decreases the level of the neutral lipid        triacylglycerol; and    -   iii) Does not change the levels of phosphatidic acid and        phosphatidylserine.

None of these effects of an anti-aging compound has been reportedbefore.

The present disclosure will be more readily understood by referring tothe following examples which are given to illustrate embodiments ratherthan to limit its scope.

Example I Identification of Previously Unknown Natural Anti-Aging PlantExtracts

The wild-type strain of the yeast S. cerevisiae BY4742 was cultured in asynthetic minimal YNB medium initially containing 2% glucose andsupplemented with 20 mg/I histidine, 30 mg/I leucine, 30 mg/I lysine and20 mg/I uracil. For assessing the longevity-extending efficiency of aplant extract, it was added at one of the following concentrations:

concentration “0”—i.e., only ethanol, a solvent used as a vehicle fordelivering various compounds into a cell, was added at a finalconcentration of 0.5%, 1.5%, 2.5% or 5%; and

concentrations 0.1%, 0.3%, 0.5% or 1% of plant extract in 0.5%, 1.5%,2.5% or 5% ethanol (final concentration), respectively.

Cells were cultured at 30° C. with shaking. A sample of cells was takenfrom a culture every day. A fraction of the sample was diluted in orderto determine the total number of cells using a haemocytometer.

Another fraction of the cell sample was diluted and serial dilutions ofcells were plated in duplicate onto plates with YP medium containing 2%glucose as carbon source. After 2 days of incubation at 30° C., thenumber of colony forming units (CFU) per plate was counted. The numberof CFU was defined as the number of viable cells in a sample. For eachculture, the percentage of viable cells was calculated as follows:(number of viable cells per ml/total number of cells per ml)×100. Thepercentage of viable cells in mid-logarithmic phase was set at 100%.

To quantitatively assess and compare the effects of various naturalproducts added at different concentrations on longevity ofchronologically aging yeast, the following two measures of chronologicallifespan were calculated: (i) mean lifespan, the number of days requiredfor a culture of yeast cells to reach 50% viability; and (ii) maximumlifespan, the number of days required for a culture of yeast cells toreach 10% viability (as show in FIG. 1). Each trial was done intriplicate and results are the average of the three trials.

A total of 31 plant extracts were selected as the one that maypotentially have anti-aging effects. To test their potential anti-agingeffects, these plant extracts were assessed for their efficiency toextend the chronological lifespan of the wild-type strain of the yeastS. cerevisiae BY4742. The table of the Example II resume the results for6 ‘positive’ extracts (i.e. the ones that significantly increase boththe mean and maximum lifespans of yeast), whereas FIG. 2 demonstratesexamples of ‘negative’ extracts. Among the 31 plant extracts tested, 6extracts were found to be positive. More than 3500 trials were performedto identify such positive extracts, to establish their most efficientlongevity-extending concentrations and to demonstrate the additiveeffects between their anti-aging effects (as discussed at Example III).

Example II Plant Extracts from Different Commercial Sources ExhibitSimilarly High Anti-Aging Effects

To be sure that the extraction process is not a limitation, each of the6 positive plant extracts was obtain from different commercial sourcesand assays was done like described in the Example I. Similar highanti-aging effects were seen with all commercial sources (Table 1).These data provide evidence that the extraction process and thecommercial source are not limiting factors for the previously unknownpotent anti-aging plant extracts described herein.

TABLE 1 Plant extracts from different commercial sources exhibitsimilarly high anti-aging effects. Different commercial sources Amean/max Plant extracts lifespan (%) B C D E F G Black Cohosh 195/100similar similar Valerian 185/87  similar similar similar similar similarsimilar Passion flower 180/80  similar similar similar similar similarsimilar Ginkgo biloba 145/104 similar similar similar similar similarsimilar Celery seed 160/107 similar similar similar White willow 475/369similar similar similar similar similar similar

Example III Beneficial Effects Between Combinations of Different PlantExtracts

Because aging is known to be modulated by several signaling pathwaysthat are controlled by different chemical compounds, the most efficientanti-aging approach is to use mixes of multiple natural products (suchas plant extracts) that are capable of modulating different signalingpathways of longevity regulation. Some of these mixes are expected toprovide improved beneficial effects when combined (i.e. mutuallyamplifying) effects on longevity. To identify the potential beneficialeffects between the different combinations of anti-aging plant extractspresented at the Example I, different pairwise combinations of theseextracts were tested.

For assessing the longevity-extending efficiency of each pair of one ofthe anti-aging plant extracts identified in Example I, each of them wasadded with another one at different concentrations. The different mixesof plant extracts were assessed for their effects on the chronologicallifespan of WT strain of yeast. The pairwise mixtures of the followingplant extracts extended the mean and maximum lifespans ofchronologically aging yeast to significantly higher extent that each ofthe plant extracts alone. Table 2 show the best values obtained.

TABLE 2 Beneficial effects between combinations of different plantextracts Alone (%) mean Formulated with (%) Plant extracts lifespan 4 56 8 12 21 Black Cohosh (4) 195 NA 365 390 155 505 850 Valerian (5) 185365 NA 385 375 510 775 Passion flower (6) 180 390 385 NA 320 365 865Ginkgo biloba (8) 145 155 375 320 NA 495 665 Celery seed (12) 160 505510 365 495 NA 570 White willow (21) 475 850 775 865 665 570 NA NA = notapplicable NTY = not tested yet

Example IV Beneficial Effects Between Combinations of Different PlantExtracts and Well-Known Anti-Aging Chemical Compounds

Currently known natural anti-aging compounds (resveratrol (Res),spermidine (Spe), metformin, and myriocin) delay aging in differentorganism, improve health and extend lifespan by targeting differentlongevity-defining cellular processes that are controlled by differentsignaling pathways. Therefore, as for Example III, it is plausible thatif two or more of these compounds when added together, or with theanti-aging plant extracts of Example I, may exhibit an increaseaging-delaying effect by enhancing the beneficial effect of each otheron health and longevity. As for Example III, to identify the possibleincreased effects between the different anti-aging plant extractspresented in Example I and the currently known natural anti-agingcompounds, their different combination was tested.

These different mixtures were assessed for their effects on thechronological lifespan of WT strain of yeast. It was observed that asynergistic anti-aging effect was noted when combining the Black Cohoshextract with Spermidine; Valerian extract with Resveratrol, metformin,myriocin and Spermidine; the Passion flower extract with Resveratrol,metformin, myriocin, and Spermidine; the Ginkgo biloba extract withSpermidine and myriocin; the Celery seed extract with Resveratrol,metformin and myriocin; and the White willow extract with Resveratrol,metformin, myriocin and Spermidine. The mixtures of the mentioned knownnatural anti-aging compounds and the newly identify anti-aging plantextracts as listed and identified hereinabove extended the mean and/ormaximum lifespans of chronologically aging yeast to significantly higherextent that each of them alone.

Example V Mass Spectrometry-Based Lipidomic Analyses of Yeast CellsFollowing Treatment with Passion Flower Extract Alone, Resveratrol Aloneor with a Mixture of Passion Flower Extract and Resveratrol

The wild-type strain of the yeast S. cerevisiae BY4742 (MATα his3Δ1leu2Δ0 lys2Δ0 ura3Δ0) was cultured in a synthetic minimal YNB medium(0.67% Yeast Nitrogen Base without amino acids) initially containing 2%glucose and supplemented with 20 mg/I histidine, 30 mg/I leucine, 30mg/I lysine and 20 mg/I uracil, as well as with a mixture of passionflower extract and resveratrol. Samples of cells were taken from aculture at days 1, 3, 6, 8 and 10 of culturing. Lipids were extractedfrom whole cells and then analyzed by quantitative mass spectrometry(see FIG. 5).

Example VI Specific Plant Extracts on Cellular Concentrations of VariousLipids in a Wild-Type Strain

As a next step towards understanding mechanisms by which the newlyidentified aging-delaying extracts extend yeast longevity, a massspectrometry (MS)-based quantitative analysis of many lipid classes wasused to investigate how the six new anti-aging plant extracts influencecellular concentrations of various lipids in a wild-type strain. As anexample the White Willow extract was found to: (1) considerably increasecellular concentrations of phosphatidic acid (PA), phosphatidylserine(PS), phosphatidylethanolamine (PE), phosphatidylcholine (PC) andphosphatidylinositol (PI); (2) greatly decrease cellular concentrationof triacylglycerols (TAG; also known as fats); and (3) have no effect oncellular concentration of cardiolipin (CL) (FIG. 6). These findingssuggest that the White Willow extract extends longevity ofchronologically aging yeast by attenuating the synthesis of TAG (themajor form of energy storage in yeast and other organisms, includinghumans) from diacylglycerol (DAG), PE and PC (FIG. 6).

Example VII Extracts Slows the Progression of Yeast Chronological Agingby Differently Modulating Certain Cellular Processes

As another step towards understanding mechanisms by which the newlyidentified aging-delaying extracts extend yeast longevity, differentcellular processes were investigated. Each of the sixlongevity-extending plant extracts is a geroprotector which delays theonset and decreases the rate of yeast chronological aging by eliciting ahormetic stress response. Accordingly, each of these extracts slows theprogression of yeast chronological aging by differently modulatingcertain cellular processes. These processes include mitochondrialrespiration, maintenance of mitochondrial membrane potential, reactiveoxygen species homeostasis, protection of cellular proteins and membranelipids from oxidative damage, stabilization of mitochondrial and nuclearDNA, cell protection from chronic oxidative and thermal stresses, andlipolytic degradation of neutral lipids deposited in lipid droplets.

FIG. 3 presents the results. PE4 (Black Cohosh), PE5 (Valerian), PE6(Passion flower), PE8 (Ginkgo biloba), PE12 (Celery seed) and PE21(White willow) delay yeast chronological aging and have differenteffects on several longevity-defining cellular processes. Arrowspointing at boxes with the terms of longevity-defining cellularprocesses denote activation of these processes, T bars denote inhibitionof these processes, whereas lines with filled circles denote change inthe age-related chronology of intracellular ROS. The thickness of sucharrows, T bars and lines with filled circles correlates with the extentto which a PE activates, inhibits or alters the age-related chronology(respectively) of a particular longevity-defining cellular process.Arrows and T bars pointing at boxes with the term “AGING” denoteacceleration or deceleration (respectively) of yeast chronologicalaging. The characterisations of these processes are clear demonstrationsof metabolic anti-aging impacts.

Example VIII Extracts Slows the Progression of Yeast Chronological Agingby Different Mechanism of Action

Aging of multicellular and unicellular eukaryotic organisms is a complexbiological phenomenon affecting many cellular processes. These numerouscellular processes are modulated by signaling pathways that areconserved across phyla and converge into a network regulating longevityin evolutionarily distant organisms. In yeast, this network includes thefollowing signaling pathways: (1) the pro-aging PKA (protein kinase A)pathway; (2) the pro-aging TORC1 (target of rapamycin complex 1)pathway; (3) the pro-aging PKH1/2 (Pkb-activating kinase homolog)pathway; (4) the anti-aging SNF1 (sucrose non-fermenting) pathway; (5)the anti-aging ATG (autophagy) pathway (FIG. 4). Moreover, SCH9 is apro-aging protein kinase stimulated by the TORC1 and PKH1/2 pathways,whereas RIM15 is an anti-aging protein kinase inhibited by the PKA andTORC1 pathways (FIG. 4).

The effects of single-gene-deletion mutations eliminating differentprotein components of pro-aging and anti-aging signaling pathways wereused to identify the mechanism of action of each anti-aging extract.This is an essential step towards understanding mechanisms by which theaging-delaying extracts extent longevity. This enable to identify alongevity-defining signaling pathway (or pathways) targeted by each ofthem.

FIG. 4 shows how PE4 (Black Cohosh), PE5 (Valerian), PE6 (Passionflower), PE8 (Ginkgo biloba), PE12 (Celery seed) and PE21 (White willow)extracts delay yeast chronological aging via the longevity-definingnetwork of signaling pathways/protein kinases. Activation arrows andinhibition bars denote pro-aging processes or anti-aging processes.

Consequently, these extracts slow aging in the following ways: 1) plantextract 4 decreases the efficiency with which the pro-aging TORC1pathway inhibits the anti-aging SNF1 pathway; 2) plant extract 5mitigates two different branches of the pro-aging PKA pathway; 3) plantextract 6 coordinates processes that are not assimilated into thenetwork of presently known signaling pathways/protein kinases; 4) plantextract 8 diminishes the inhibitory action of PKA on SNF1; 5) plantextract 12 intensifies the anti-aging protein kinase Rim15, and 6) plantextract 21 inhibits a form of the pro-aging protein kinase Sch9 that isactivated by the pro-aging PKH1/2 pathway (FIG. 4).

Example IX Synergy of the Six Extracts Together

Each of the six plant extracts (PEs) delays aging through differentsignaling pathways and/or protein kinases (FIG. 4, Example VIII).Therefore, this is why at the Example III, when these PEs are mixed invarious combinations, some of the combinations display additive orsynergistic effects on the aging-delaying efficiencies of each other. Tosee at which extend it's possible to increase the anti-aging impact, atrial was done in the same condition as in Example I, but at 0.1% ofeach extract. All together, these 6 extracts were able to extant themean lifespan by 763%, which is better than any combination shown atExample III (Table 2).

Example X Extracts Slows Aging Process and Improve Healthy Aging inNematodes

Aging of multicellular eukaryotic organisms is a complex biologicalphenomenon affecting many cellular processes. Bristol N2 worms were usedto test the impact of 2 extracts. Culture and handling of nematodes wereconducted as previously described (Brenner et al., 1974, Genetics, 77:71-94). Worms were maintained at 20° for all the experiments.

For lifespan assays, worms were synchronized by the alkalinehypochlorate method (Porta-de-la-Riva et al., 2012, J Vis Exp., 64:e4019). Synchronized L1 larvae were grown on bacteria to L4 larvae. Atthis stage, worms were seeded to plates containing the compound extractsor vehicle, which was considered day 1 of the experiment. Animals wereconsidered dead when they ceased moving or responding to prodding.Animals that crawled off the plates, “exploded”, or had a visible eglphenotype were discarded from the lifespan analysis.

Extract PE5 (Valerian) and PE8 (Ginkgo biloba) were tested and shownpositive impact on longevity and vitality of nematodes. The nematodeswere more active during the aging process. Extracts PE5 showed positiveresults at all the tested concentration (50, 100 and 250 ug/ml) and PE8at the two most important concentrations (100 and 250 ug/ml).

Example XI Extracts Slows Aging in Animal Accelerate Aging Model ofWerner Syndrome

Werner syndrome (WS) is a human autosomal recessive disordercharacterized by genomic instability, the premature aging and the onsetof a number of age-related diseases. The defective enzyme responsiblefor WS possesses a 3′-5′ exonuclease activity in addition to a 3′-5′ DNAhelicase activity and is involved in DNA repair, replication,transcription, and telomere maintenance. A mouse model was used with adeletion in the helicase domain of the murine WRN homologue thatrecapitulates many of the WS phenotypes, to test the anti-aging impactsof a mixture of plant extracts (a progeria model). The six anti-agingplant extracts described in this invention was added in animal waterwith resveratrol and olive polyphenols during a 12 months study.

After 12 months, animals were about 2 month younger considering theirbody weight. Glucose metabolism (fasting blood glucose and insulin,OGTT) was improved and kept as it was for 8 weeks animals. These tworesults, body weight and glucose metabolism, are related to a betterenergy usage regulation as seen for each extract as mitochondrialbenefits. Their muscle resistance was greatly increased and underscoresa possible utilization of these extracts in sarcopenia to conservemuscle during the aging process. This muscle impact is also related to abetter energy metabolism and mitochondrial function during the agingprocess.

The memory was also slightly improved (maze test).

While the disclosure has been described in connection with specificembodiments thereof, it will be understood that it is capable of furthermodifications and this application is intended to cover any variations,uses, or adaptations, as may be applied to the essential featureshereinbefore set forth, and as follows in the scope of the appendedclaims.

1. An anti-aging composition comprising at least two different anti-aging agents, wherein said at least two anti-aging agents are two different plant extracts or one plant extract and a second anti-aging agent, wherein the plant extract is at least one of a Black cohosh extract, a Valerian extract, a Ginkgo biloba extract, a Celery seed extract, a White willow extract and a Passion flower extract, and wherein the at least two anti-aging agents have an superior effect on regulating longevity compared to one anti-aging agent.
 2. The composition of claim 1, wherein the second anti-aging agent is resveratrol, metformin, myriocin, or spermidine.
 3. The composition of claim 1 or 2, wherein the at least two anti-aging agents modulate at least two pathways that regulate longevity.
 4. The composition of claim 3, wherein said pathways are TORC1, cAMP/PKA, PKH1/2, SNF1/AMPK or ATG pathways.
 5. An anti-aging composition comprising at least one plant extract selected from the group consisting of a Black cohosh extract, a Valerian extract, a Ginkgo biloba extract, a Celery seed extract, a White willow extract and a Passion flower extract, and a carrier.
 6. The anti-aging composition of claim 5, said composition comprising a Black cohosh extract, a Valerian extract, a Ginkgo biloba extract, a Celery seed extract, a White willow extract and a Passion flower extract.
 7. The anti-aging composition of claim 5, said composition further comprising resveratrol, metformin, myriocin, or spermidine.
 8. The anti-aging composition of claim 7, comprising a Black Cohosh extract and Spermidine
 9. The anti-aging composition of claim 7, comprising a Valerian extract with Resveratrol, metformin, myriocin or Spermidine.
 10. The anti-aging composition of claim 7, comprising a passion flower extract and resveratrol, metformin, myriocin, or spermidine.
 11. The anti-aging composition of claim 7, comprising a Ginkgo biloba extract and myriocin or spermidine.
 12. The anti-aging composition of claim 7, comprising a Celery seed extract and Resveratrol, metformin or myriocin.
 13. The anti-aging composition of claim 7, comprising a White willow extract and Resveratrol, metformin, myriocin or Spermidine.
 14. The anti-aging composition of claim 5, wherein said composition delays the onset and progression of age-related disorders.
 15. The anti-aging composition of claim 14, wherein said age-related disorders are cardiovascular disorders, glycemic disorders, neurodegenerative disorders or osteoporosis disorders.
 16. The anti-aging composition of claim 15, formulated as a cosmetic composition, a dermatological composition, a nutraceutical composition or a pharmaceutical composition. 17.-18. (canceled)
 19. A method of prolonging longevity of a subject comprising administering to said subject an effective amount of the composition of claim 1 or
 5. 20. The method of claim 19, wherein the subject is a human, a mice or a nematode. 